WO1998050353A1

WO1998050353A1 - Vitamin d3 derivatives and process for producing the same

Info

Publication number: WO1998050353A1
Application number: PCT/JP1998/001979
Authority: WO
Inventors: Hiroaki Takayama; Katsuhiro Konno; Toshie Fujishima
Original assignee: Teijin Limited
Priority date: 1997-05-02
Filing date: 1998-04-30
Publication date: 1998-11-12
Also published as: EP0957088B1; EP0957088A4; DE69810273T2; JP3923090B2; US6982257B1; EP0957088A1; ATE229937T1; DE69810273D1

Abstract

1,25-Dihydroxy-2-methyl vitamin D3 derivatives represented by general formula (I) wherein R1 and R2 each independently represents hydrogen or tri(C1-7alkyl)silyl; and the asymmetric carbon atoms at the 1-, 2- and 3-positions each independently has an α- or β-configuration. These compounds are useful as remedies for osteoporosis, rachitis, accessory thyroidal hyperenergia, etc.

Description

Bright fine manual vitamin 13 ₃ derivatives and their preparation art

The present invention relates to a novel Pitamin 13 ₃ derivative AND ITS preparation having a methyl group at the 2-position. More particularly, 1 useful as a osteoporosis therapeutic agents, 2 5 - relates methyl vitamin D ₃ derivatives and their preparation - dihydric Dorokishi 2. Background art

As material activated vitamin 13 ₃ to control the metabolism of calcium Ya-phosphate salt in vivo, to the very important role it is widely recognized throughout the scientific literature patent publications and general ever. It is also a well-known fact that various vitamin D derivatives are used as therapeutic agents for dysbimin D metabolism, including osteoporosis and rickets.

Furthermore, various biological activities observed in calcium regulation effects or other vitamin D ₃ vitamin D receptions binding affinity and vitamin binding affinity of various action selectivity expressed by differences in the D-binding protein evening to single Some have interpreted that it is due to

Known 2-substituted vitamin 0 ₃ derivative, 1-hydroxyl group is α-coordinated, 2-position i3 coordinating substituents (unsubstituted or terminally substituted with a hydroxyl group (: E - the C ₆ . dihydroxyvitamin D ₃ derivative - a linear alkyl group, terminally linear Arukiruokishi group E one C ₆ to Flip substituted with a hydroxyl group, 1, 2 5 with E one C ₅ of alkenyl group or a hydroxyl group) (Kobayashi et al., Japanese Pharmaceutical Society 1st Annual Meeting (1996), Abstracts of Lectures 3, p88). Also, the hydroxyl group at the 1-position is a para-coordinate and the 2-position has a tri-coordinate substituent (3-hydroxypropyl or 3-fluoropropyl). 1,25-dihydroxyvitamin D _Three derivatives are known (Posner, H., J. Org. Chem., 1995, 60, 46 17).

In addition, studies have been reported on other stereoisomers of the 1, 25-dihydroxy-12-methylvitamin D ₃ derivative with respect to its asymmetric carbons at the 1, 2, and 3 positions. The Pharmaceutical Society of Japan 1st 16th Annual Meeting (1996), Abstracts of Lectures 2, p9).

However, among the 1, 25-dihydroxy-12-methylvitamin D ₃ derivatives, no stereoisomer (2OS isomer) different from the natural one is known for the configuration at the carbon atom at position 20. . Therefore, the configuration of the carbon atom at position 20 may affect the binding affinity to the vitamin D receptor, the binding affinity to the vitamin D binding protein. Not known until now.

Incidentally, 2-substituted vitamin 0 ₃ derivatives of preparation also mosquito ^ either position 1 shown in the above document, position 2, and 3 of the stereoisomers of the chiral carbon, of a particular combination It is only capable of producing isomers, and does not show a method for efficiently producing an arbitrary combination of isomers.

In recent years, the following general formula (Π ')

[In the formula, X represents a bromine atom or an iodine atom. An exomethylene compound represented by the following general formula (1 ′)

[Wherein, R ₃ and R ₄ each independently represent a hydrogen atom or a tri (C 1 -C ₇ hydrocarbon) silyl group. ]

Active bi evening novel method of synthesizing Mi emissions 0 ₃ by reacting in represented by Enin compounds were published (Trost, BM;... J. Am Chem Soc, 1992, _Π 4, 9836) . However, there is no known example using an enyne compound having a substituent such as a methyl group at the 4-position. Disclosure of the invention

An object of the present invention relates to novel 1, 2 5-dihydroxy sheet having biological activity - in the child provides a 2-Mechirubi evening Min D ₃ derivatives and their preparation.

According to the present invention, the above object of the present invention is achieved, first, by the following general formula (I)

Wherein R i and R ₂ are each independently a hydrogen atom or a tri

(C ^ —C ₇ alkyl) represents a silyl group. Here, the configurations of the asymmetric carbons at the 1-, 2-, and 3-positions are each independently para- or β-coordinate. ]

This is achieved by a 1,25-dihydroxy-2-methylvitamin D ₃ derivative represented by

That is, in the vitamin D ₃ derivative of the present invention, the configuration at the 1, 2, and 3 positions is respectively

(1) Combination of α-coordinate, para-coordinate and α-coordinate

(2) Combination of α-coordinate, H-coordinate and j3-coordinate

(3) Combination of coordination, / 3 coordination and α coordination

(4) Combination of α-coordinate, / 3-coordinate,) 3-coordinate

(5)) 3-coordinate, paracoordinate, combination of paracoordinates

(6) Combination of 0-coordinate, α-coordinate, and] 3-coordinate

(7) Combination of j8 coordination,) 3 coordination and paracoordination

(8) / 3 coordination, / 3 coordination,) Combination of 3 coordination

All eight types are included. Furthermore, a mixture containing a plurality of any of these eight stereoisomers at an arbitrary ratio is also included in the scope of the present invention. The notation for the configuration of vitamin D is a convention, that is, the `` coordinate '' used at the 1st, 2nd, and 3rd positions means the bond from the top of the paper, ] 3 Coordination "means binding from below the paper.

According to the present invention, the object of the present invention is:

It is achieved by vitamin D ₃ derivatives of the preparation of formula (I). That is, the following general formula (II)

[In the formula, X represents a bromine atom or an iodine atom. ]

An exomethylene compound represented by the following general formula (ΙΠ)

[Wherein, R ₃ and R ₄ each independently represent a hydrogen atom or a tri (C i -C hydrocarbon) silyl group. ]

Is reacted with an enyne compound represented by the formula (I) in the presence of a palladium catalyst to remove the tri-C ₇ hydrocarbon) silyl group, if necessary, to obtain a compound represented by the formula (I) 2 is a 5-dihydrazide Dorokishi one 2 process for preparing methyl vitamin 0 ₃ derivatives. BEST MODE FOR CARRYING OUT THE INVENTION

In the present invention, the term “tri-c ₇ alkyl) silyl group means a silyl group substituted by three independent linear or branched C 1 -C ₇ alkyl groups, and among them, Trimethylsilyl, triethylsilyl, or t-butyldimethylsilyl is preferred.

Preferable specific examples of the 1,25-dihydroxy-2-methylbiamine D ₃ derivative represented by the above formula (I) include:

(20 S) 11, 25-dihydroxy 1 2) 3-methyl-3) 3-bimin D ₃ …… (68)

(2 OS) — 1) 3, 2 5 —Dihydroxy 2) 3 —Methyl- 3) 3—Bimin D ₃ …… (69)

(2 0 S) 1 1, 2 5-dihydroxy-2 i3-methyl-3-hibi Yumin D ₃ ...... (70)

(2 0 S) 1/3, 25 5 -dihydroxy 2 j3 -methyl-3 α-bimin D ₃ …… (7 1)

(2 OS) — 1 α, 25 — Dihydroxy 1 2 methyl 3) 3 — Bimin D ₃ …… (7 2)

(2 0 S) 1 1) 3, 2 5 —dihydroxy 2 2 methyl 3 3 — vitamin D ₃ …… (73)

(2 0 S)-l, 2 5 —Dihydroxy 1 2 1 Methyl 3 ο; —Bimin D ₃ …… (7 4)

(2 0 S) — 1) 3, 2 5 —dihydroxy 2 2 1 methyl 3 3 hibi Yumin D ₃ …… (7 5)

(2 0 S) — 1, 2 5 — dihydroxyl 2 ι3 — methyl-1 3 | 3 — bimin D ₃ — 1, 3 — bis (trimethylsilyl) ether …… (7 6) (2 OS) — l j3, 25 — dihydroxyl 2; 3 — methyl-3 / 3 — bi-min D ₃ — 1, 3 — bis (trimethylsilyl) ether …… (7

7)

(2 0 S) one 1 shed, 2 5 - dihydric Dorokishi 2/3 - methyl one 3 Hibi evening Min D ₃ - 1, 3 _ bis (trimethylsilyl) ether .... (7

8)

(2 0 S) one 1/3, 2 5 - dihydric Dorokishi 2 0 - methyl one 3 Hibi evening Min D ₃ - 1, 3 - bis (trimethylsilyl) ether .... (7

9)

(2 0 S) - 1 α , 2 5 - dihydric de Roxy 2 Fei one methyl - 3 i3 - bi evening Min D ₃ - 1, ₃ - bis (Bok trimethylsilyl) ether .... (8 0)

(2 0 S) one 1 beta 2 5 - dihydric Dorokishi - 2 alpha - methyl - 3 j3 - bi evening Min D ₃ - 1, ₃ - bis (trimethylsilyl) ether ...... (8 1)

(2 0 S) one 1 shed, 2 5 - dihydroxy over 2 alpha - methyl one 3 Hibi evening Min D ₃ _ l, 3 - bis (trimethylsilyl) ether ...... (82)

(2 0 S) one 1) 3, 2 5 - dihydric Dorokishi - 2 alpha - methyl - 3 Fei - Pi evening Min D ₃ - 1, 3 - bis (trimethylsilyl) ether ...... (8 3)

(2 0 S) - 1 α , 2 5 - dihydroxy one 2 i3 - methyl one 3) 3 - bi evening Min D ₃ _ l, 3 - bis (t - heptyl dimethylsilyl) ether ...... (8 4)

(2 0 S) one 1/3, 2 5 - dihydric Dorokishi - 2/3 - methyl-3/3 - bi evening Min D ₃ - 1, 3 - bis (t - heptyl dimethylsilyl) ether ...... (8 5) (2 0 S) one 1 shed, 2 5 dihydric Dorokishi - methyl one 3 alpha-vitamin 1 ₃ - 1, 3-bis (t one heptyl dimethylsilyl) ether ...... (8 6)

(20 S) — l i3, 25-dihydroxy-1/3-methyl _ 3 hibi Yubimin D ₃ _ l, 3-bis (t-butyldimethylsilyl) ether …… (87)

(2 0 S) - 1 α , 2 5- dihydric Dorokishi one second non-methyl-3 JS-bi evening Min D ₃ - 1, 3- bis (t one heptyl dimethylsilyl) E one ether ... (8 8)

(20 S) — 1/3, 25-dihydroxy- 1 2α-methyl- 1 3 i3-bi-umin D ₃ — 1,3-bis (t-butyldimethylsilyl) ether …… (89)

(2 0 S) one 1 shed, 2 5- dihydric Dorokishi 2 Facial Mechiru one 3 Facial velvetleaf evening Min D ₃ - 1, 3- bis (t - butyldimethylsilyl) E one ether ... (9 0)

(2 0 S) one 1) 3, 2 5 dihydric Dorokishi one 2 alpha-methyl one 3 alpha-bi Tami D ₃ - 1, 3- bis (t chromatography butyldimethylsilyl) ether ...... (9 1)

And the like.

In addition, in the method for producing the vitamin D ₃ derivative represented by the above formula (I), the en-yne compound represented by the above formula (III), which is a starting material, is substituted by its 3-, 4-, and 5-positions. It may be all stereoisomers derived from asymmetric carbon, or a mixture of these in any proportion, but their configuration is conserved during the reaction, and the 1,2,5-diphenyl has the corresponding configuration. Dorokishi 2 _ Mechirubi evening Min D ₃ derivative is generated. The palladium catalyst used in the present production method is a combination of a zero-valent or divalent organic palladium compound and a trisubstituted phosphorus compound. As such an organic palladium compound, tetrakis (triphenyl) Nylphosphine) palladium, tris (dibenzylideneacetate palladium), tris (dibenzylideneacetone) palladium chromate form, palladium acetate, etc. Examples of the trisubstituted phosphorus compound include triphenylphosphine, Tributyl phosphine, etc. As the palladium catalyst combining both, tris (dibenzylideneacetone) palladium and triphenylphosphine, tris (dibenzylideneacetone) palladium chloroform and triphenylphosphine are preferable. The mixing ratio is preferably 1: 1 to 1:10.

Here, the molar ratio of the exomethylene compound represented by the above formula (II) to the enyne compound represented by the above formula (III) is desirably in the range of 1: 5-5: 1. The palladium catalyst is used in an amount of 0.1 to 100 mol%, preferably 1 to 20 mol%, based on the exomethylene compound.

In the reaction between the exomethylene compound represented by the above formula (II) and the enyne compound represented by the above formula (III), the reaction solvent is a non-polar solvent such as hexane, heptane or toluene, getyl ether, Examples thereof include polar solvents such as tetrahydrofuran, dioxane, dimethoxetane, N, N-dimethylformamide, and acetonitrile, and a mixed solvent thereof. Of these, heptane and toluene are preferred. Furthermore, when these solvents are used in the reaction, it is desirable to carry out a treatment such as distillation or nitrogen substitution in advance. The reaction is carried out at a temperature ranging from room temperature to the boiling point of the solvent.

Further, in order to capture an acid such as hydrogen halide generated in the reaction system, it is preferable to add a base such as triethylamine, diisopropylethylamine or the like to carry out the reaction. As the amount of the base to be added, it is preferable to use at least one equivalent of the reactant represented by the above formula (II) or the above formula (III), which is used in excess. Of the vitamin D ₃ derivatives of the above formula (I) obtained by the above reaction and those in which R ₂ represents a tri (di-C ₇ alkyl) silyl group, a deprotection reaction may be carried out if necessary. , And R ₂ can be converted to those of a hydrogen atom.

Such a deprotection reaction can be performed according to a known method (for example, Calveley, M. J .; Tetrahedron, 20, 4609, 1987, Ho, PT .; Tetrahedron Letters, 1623, 1978). In this case, examples of the deprotecting agent include tetrabutylammonium fluoride, lithium tetrafluoroporate, pyridium-P-troen sulfonate and camphorsulfonic acid.

The exomethylene compound represented by the above formula (II) is synthesized according to a known method (B. Fernandez et al., J. Org. Chem., 1992, 57, 3173; J. Calverley et al., Chem. Lett., 1993, 3, 1845, A. Kutner et al., J. Org. Chem., 1988, 53, 3450).

Preferred specific examples of the enepine compound represented by the above formula (ΙίΙ) used in the production method of the present invention include:

(3 R, 4 R, 5 R) 1, 3, 5 —dihydroxy— 4—methyl 1 —octene 7 —in …… (2 2)

(3 S, 4 R, 5 R) 1, 3, 5-dihydroxy-4-methyl-1-octene 1 7-in ... (2 3)

(3 R, 4 R, 5 S) 1, 3, 5 — dihydroxy 4 — methyl 1 — octene 1 7 — …… (2 4)

(3 S, 4 R, 5 S) 1, 3, 5 — dihydroxy _ 4 — methyl 1 — octene 1 7 — in …… (25)

(3 R, 4 S, 5 R) — 3, 5 —Dihydroxy— 4 —Methyl-1 1-octene 7 —In …… (26)

(3 S, 4 S, 5 R) 1, 3, 5 — dihydroxy 4-methyl 1 — octene 1 7 — …… (2 7) (3 R, 4 S _: 5 S) -3,5-dihydroxy-4-monomethyloctene

(3 S, 4 S: 5 S) -3,5-Dihydroxy-4-methyl-1-octene …… (29)

(3R, 4R5R) -3,5-bis (trimethylsilyloxy) -4-methyl 1-octene-7-in…-(30)

(3S, 4R5R) -3,5-bis (tri-J-methylsilyloxy) -4-methyl-1-octen-1-in …… (3 1)

(3R, 4R.5S) -3,5-bis (tri ') methylsilyloxy) — 4-methyl-1-octene-1-in… (32)

(3S, 4R.5S)-3, 5-bis (trimethylsilyloxy)-4-methyl 1-octene-7-in… (33)

(3R, 4S.5R) -3,5-bis (tri ') methylsilyloxy) 1 4-methyl 1-octene 7 1-in… (3 4)

(3 S, 4 S. 5 R) — 3, 5-Bis (tri-J-methylsilyloxy)-4-methyl 1 —octen-1-in… (35)

(3R, 4S.5S) -3,5-Bis (trimethylsilyloxy) _4-methyl 1-octene-7-in… (36)

(3 S, 4 S. 5 S) .- 3,5 —bis

One 4-one methyl 1-octene—7 one-in… (3 7)

(3R, 4R.5R) -3,5—bis (t-butyldimethylsilyloxy) -14-methyl-1-octene-7-yne …… (38) (3S, 4R.5R)- 3, 5-bis (t-butyldimethylsilyl.)-4 methyl-1—octen-7-in …… (39)

(3R, 4R.5S) — 3,5-bis (t-butyldimethylsilyloxy) -14-methyl-1-octene-toin …… (40)

(3S, 4R.5S) -3,5-bis (t-butyldimethylsilyloxy) -14-methyl-1-octene-7-yne …… (41) (3R, 4S, 5R) 1,3,5-bis (t-butyldimethylsilyloxy) —4-methyl-1-octene-7—in …… (4 2)

(3 S, 4 S, 5 R) — 3, 5 _ bis ((; 1-butyldimethylsilyloxy) — 4-methyl-1- 1-octene 7 —in …… (4 3) (3 shaku, 4 3, 5 3 ) — 3, 5 — Bis (t-butyldimethylsilyloxy) — 4-methyl-1 —octene — 7 _in …… (4 4)

(3S, 4S, 5S) -1,3,5-bis (t-butyldimethylsilyloxy) —4-methyl-1-octen-17-yne... (45) and the like.

The chain compound represented by the above formula (UI) used in the production method of the present invention can be synthesized, for example, according to the following scheme 1.

Oxidation OHC.wittig reaction

, OR, epoxidation, Y,. R,

(V I I (V I I I)

Scheme 1 In the above scheme 1, represents a tri-C ₇ alkyl) silyl group or a (^ -C ₇ alkyl) di ((^-^^ aryl) silyl group. Preferred examples thereof include a trimethylsilyl group and a triethylsilyl group. R ₂ represents a protecting group which forms an acetal together with the oxygen atom to be bonded, and a methoxymethyl group, a methoxyxetoxymethyl group, and t-butyldimethylsilyl group, and t-butyldiphenylsilyl group. And a tetrahydrobiranyl group are preferred.

This manufacturing method can be performed as follows. That is, the hydroxyl group of a commercially available optically active ester compound (IV) is silyl protected in the presence of a base to obtain a compound (V). As the silylating agent, triethylsilyl chloride, t-butyldimethylsilyl chloride, t-butyldiphenylsilyl chloride, triethylsilyl triflate, t-butyldimethylsilyl triflate and the like are preferably used. As the base, a usual base such as triethylamine, 2,6-lutidine or imidazole is used.

Next, the compound (V) is reduced with a hydride reducing agent to obtain the alcohol (VI). As the hydride reducing agent, lithium aluminum hydride, diisobutyl aluminum hydride, and the like are preferable. Further, the generated hydroxyl group is oxidized with dimethyl sulfoxide / oxalyl chloride TPAP (tetrapropylammonium pentalate) ZN-methylmorpholine-N-oxide or the like to obtain an aldehyde (VII). The reaction is carried out to obtain a methylene compound (VIII).

Next, the double bond is converted to an epoxide compound (IX) using a peroxide reagent such as hydrogen peroxide or metachloroperbenzoic acid, and then reacted with an acetylene derivative shown in the scheme in the presence of a base such as alkyllithium. Compound (X) is obtained. Compound (X) is produced as a mixture (1: 1) of two diastereomers based on the stereoisomerism of the hydroxyl group, which can be easily separated and purified by ordinary separation operations such as column chromatography. Wear. In addition, the configuration of the hydroxyl group of the separated diastereomer can be determined by measuring the MTPA ester of (R) — and (S) — and measuring i HNM R (Kusumi et al., Organic Synthesis Journal of the Chemical Society of Japan, 1996, 51, 462).

Further, by subjecting the separated compound (X) to the following reaction, the desired en-yne compound represented by the above formula (III) can be produced optically pure. That is, the compound

The compound (XI) is obtained by protecting the hydroxyl group of (X) with acetal. As the acetalizing agent, methoxymethyl chloride, methoxetoxymethyl chloride, dihydroxypyran and the like are used. Then, after desilylation with a fluoride reagent such as tetrabutylammonium fluoride to give a compound (II), the generated primary hydroxyl group is converted to dimethyl sulfoxide / oxalyl chloride {TPA} (tetrapropylammonium pentapentenate). -Methylmorpholine- Oxidized with N-oxide etc. to form aldehyde (XIII). Further, the aldehyde group is reacted with a vinyl Grignard reagent to obtain a compound (XIV). Finally, the desired enyne compound (III) can be obtained by removing the acetal protecting group at the 5-position hydroxyl group under acidic conditions.

This compound (III) is formed as a mixture (1: 1) of two diastereomers based on the stereoisomerism of the hydroxyl group at the 3-position, which can be easily separated and purified by ordinary separation operations such as column chromatography. You. In addition, the configuration of the hydroxyl group of the separated diastereomer can be determined by converting each of them into an acetonide formed by a hydroxyl group at the 3- or 5-position and measuring ¹³ C NMR (Rychnovsky, SD; J. Org. Chem., 1993, 58, 3511). Furthermore, it can lead to a protected silyl group at the 3- or 5-position hydroxyl group, if necessary. Examples of the silylating agent used herein include trimethylsilyl chloride, triethylsilyl chloride, t-butyldimethylsilyl chloride, t-butyldiphenylsilyl chloride, triethylsilyl triflate, and t-butyldimethylsilyl triflate. A flat base or the like is preferably used, and a normal base such as triethylamine, 2,6-lutidine or imidazole is used as the base. As the reaction conditions such as the solvent and the reaction temperature in each reaction step of the above reaction formula, conditions usually used for each reaction are applied.

In this production method, the steric configuration of the 4-position methyl group of the desired en-yne compound (III) is derived from the optically active ester compound (IV) used as the starting material, and in this synthesis route, This configuration is maintained throughout the reaction. Namely, optically active S. ether compound (IV) used as the starting material, by employing consistent reaction to maintain its configuration, key intermediates of vitamin D ₃ compounds synthesized (III) optically pure Can be manufactured.

As an example of the production of an optically pure enyne compound by this method, (3R, 4R, 5R) -13,5-bis (t-butyldimethylsilyloxy) _4-methyl-1-octene-7-yne ( The synthesis of (3 8) and (3 S, 4 R, 5 R) -1,5-bis (t-butyldimethylsilyloxy) —4-methyl-1-octen-7-yne (39) is shown in Scheme 2 below. And Scheme 3.

TBDPSC1

MeOtsu C Imidazole Me0 ₂ C

OH "OTBDPS

(46) (47) (48)

TPAP Ph ₃ P + CH ₃ Br- NMO OHC BuLi mCPBA

OTBDPS

(49) (50)

(R) -MTPA ester (54) (R) -MTPA ester (56) (S) -MTPA ester (55) (S) -MTPA ester (57)

Scheme 2

(53) (58) (59)

(60) (61)

(22) (23)

TBSOTf TBSOTf

2,6-Lutidine 2,6-Lutidine

(38) (39)

Scheme 3 [In the above scheme, TBDPSC 1 is t-butyldiphenylsilyl chloride, DIBAL-H is diisobutylaluminum hydride, TPAP is tetrapropylammonium pentalutenate, NMO is N-methylmorpholine-N-oxide, mC PBA Is metaclo-perbenzoic acid, MT PAC 1 is hymethoxy-hyr — (trifluoromethyl) phenylacetyl chloride, DMAP is 4-dimethylaminopyridine, DHP is dihydroxypyran, T sOH is tosylic acid, TBAF stands for tetrabutylammonium fluoride, TBS OTf stands for t-butyldimethylsilyl triflate, TBDPS stands for t-butyldiphenylsilyl group, TBS stands for t-butyldimethylsilyl group, and THP stands for tetrahydropyranyl group. Represents ]

In addition to this example, for example, the (4R) series can be prepared by a similar production method using the compound (52) obtained in the above scheme 2, and the (4S) series can be used as a starting material in the following optically active ester compound ( It can be synthesized by a similar production method using 6).

(64)

Example

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to these Examples.

First, a method for preparing the compound of the formula (III), which is a synthetic intermediate of the compound of the present invention, will be described as a reference example.

[Reference Example 1]

Synthesis of methyl- (S) -3- (t-butyldiphenylsilyloxy) -l-methylpropionate (47)

MeO C,

2, OTBDPS

(46) (47)

Under an argon atmosphere, methyl (S) — 3 — hydroxy 2 — methylpropionate (46) (1.9 ml, 2. Og, 16.9 mmo1) was added to 10 O ml of dichloromethane. After dissolving, imidazole (2.3 g, 32.5 mmol) and TBDPSCl (4.3 ml, 16.9 mmol) were added, and the mixture was stirred for 5 minutes. H ₂ 〇 was added, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with saturated saline, dried over magnesium sulfate, and the solvent was distilled off. The crude product was purified by column chromatography (60 g, 2% AcOEt-hexane) to obtain (47) (6.5 g, quant) as a colorless oil.

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

1.04 (9Η, s), 1.15 (3Η, d, J = 7.OHz), 2.72 (1H, dquin, J = 5.8, 7.0Hz), 3.67 (3H, s), 3.73 (1H, dd, J = 6.4, 9.8Hz), 3.83 (1H, dd, J = 9.8, 6.4Hz), 7.35-7.44 (6H, m), 7.64-7.68 (4H, m)

Sm / z325 ( ⁺ -Me- e), 299 (M -tBu) [Reference Example 2]

Synthesis of (R) -3- (t-butyldiphenylsilyloxy) 1-2-methylpropanol (48)

(47) (48)

Under an argon atmosphere, methyl (S) — 3 — (t-butyldiphenylsilyloxy) — 2 — methylpropionate (47) (1.0 g, 2.7 mmo 1) in 50 ml of dry toluene And 1 MDIBAL-H / hexane (5.7 ml, 5.7 mmol) was added at 0 ° C, and the mixture was stirred for 15 minutes, returned to room temperature, and stirred for 45 minutes. Ethyl acetate was added to the reaction solution to decompose excess DIBAL-H, and the reaction solution was extracted with 0.5 N HC1. The ethyl acetate layer was washed with saturated saline, dried over magnesium sulfate, and the solvent was distilled off. The crude product was purified by column chromatography (30 g, 4-10% AcOEt-hexane) to obtain (48) (966 mg, quatnt) as a colorless oil.

1 recitation R (400MHz, CDCI3 / TMS) 6:

0.83 (3Η, d, J-7.0Hz), 1.06 (9H, s), 1.99 (1H, ddddq, J = 4.6, 5.2, 6.2, 7.0Hz), 2.58 (1H, bs), 3.60 (1H, dd, J = 7.6, 10.1Hz), 3.97 (2H, d, J = 6.4Hz), 3.72 (1H, dd, J = 4.6, 10.1Hz), 7.37-7.46 (6H, m), 7.67-7 . 69 (4H, m)

MSm / z 328 (M ⁺ ), 271 ( ⁺ -tBu) [Reference Example 3] Synthesis of (S) 13- (t-butyldiphenylsilyloxy) -12-methylpropanal- (49)

HO ^^^ OTBDPS ^ ^JH ^{^^ 0TBDPS}

(48) (49)

Under an argon atmosphere, (R) — 3 — (t — butyldiphenylsilyloxy) 1-2 — methylpropanol (48) (725 mg, 2.2 mmo 1) was dissolved in 40 ml of dry dichloromethane. Add MS-4A (30 mg), NMO (862 mg, 11.1 mmo1) and TPAP (cat) at 0 ° C, stir for 15 minutes, return to room temperature and overnight Stirred. Then the H ₂ 0 was added, followed by extraction with acetic acid Echiru. This was washed with an ethyl acetate saturated brine, dried over magnesium sulfate, and the solvent was distilled off. The crude product was purified by column chromatography (21 g, 4% AcOEt-hexane) to give (49) (700 mg, 97%) as a colorless oil.

'ΗBrain R (400MHz, CDCI3 / TMS) δ:

1.04 (9Η, s), 1.10 (3Η, d, J = 7.0Hz), 2.56 (1H, ddddq, J = l. 3, 4.8, 6, 1, 7.0Hz), 3.87 (2H, ddd, J = 4.8, 6.1, 10.0Hz), 7.36-7.46 (6H, m), 7.63-7.67 (4H, m), 9.77 (1H, d, J = l. 5Hz)

MSm / z325 (M ⁺ -H), 269 (M ⁺ -tBu) [Reference Example 4]

Synthesis of (S) -4- (t-Butyldiphenylsilyloxy) -3'-methyl-111-butene (50) ° ^HC "^ OTBDPS ^ ^ ^ OTBDPS (49) (50)

Under an argon atmosphere, Ph ₃ P + CH ₃ Br— (2.2 g, 7.4 mmol) was suspended in 15 ml of THF, and butyllithium (5.2 ml, 9.3 mm) was suspended at 0 ° C. o 1) was added and stirred for 20 minutes. This was added to a solution of (S) —3— (t-butyldiphenylsilyloxy) -12-methylpropanal (49) (1.2 g, 3.7 mmo 1) in THF (15 ml). The mixture was added at 0 ° C, stirred for 15 minutes, returned to room temperature, and stirred for 45 minutes. A saturated aqueous ammonium chloride solution was added, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with brine, dried over magnesium sulfate, and the solvent was distilled off. The crude product was purified by column chromatography (12 g, 2% AcOEt_hexane) to obtain (50) (1.1 g, 92%) as a colorless oil.

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

1.03 (3Η, d, J = 7.0Hz), 1.05 (9H, s), 2.39 (1H, ddq, J = 6.0, 6. 7, 7.OHz), 3.49 (1H, dd, J = 6. 7, 9.7 Hz), 3.57 (1H, dd, J = 6.1, 9.7 Hz), 5.01 (3H, m), 7.35-7.44 (6H, m), 7 65-7. 68 (4H, m), 9.77 (1H, d, J = l. 5Hz) Sm / z 267 ( ⁺ -tBu)

[Reference Example 5]

Synthesis of (3 S) 1-4 1 (t-butyldiphenylsilyloxy) 1-3-methyl-1 1-butenoxide (51)

Under argon atmosphere, (50) (1.0 g, 3.1 mm o 1) was dissolved in 25 ml of dry dichloromethane, and mC PBA (1.4 g, 7.4 mm o 1) was added at 0 C. Stirred for 15 minutes. This was returned to room temperature and further stirred overnight. H ₂加え was added, and the mixture was extracted with ethyl acetate. The acetate Echiru layer was washed with saturated brine, and purified the crude product was dried over magnesium sulfate by column chromatography (3 0 g, key San 2% E t ₂ 〇- to), a colorless oil (5 1) (1. lg, quant) was obtained.

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

0.99 (3Η, d, J-6.8Hz), 1.05 (5H, s), 1.07 (4H, s), 1.58 (1H, dtq, J = 5.

0, 6.7, 7.0Hz), 2.57 (4 / 9H, m), 2.60 (8 / 9H, dd, J = 2.7, 5.0Hz), 2.

73 (4 / 9H, dd, J = 4.3, 5.0Hz), 2.76 (5 / 9H, dd, J = 4.3, 5.0Hz), 2.85 (5

/ 9H, ddd, J = 2.7, 4.3, 7.0Hz), 2.97 (4 / 9H, ddd, J = 2.7, 4, 3, 7.0Hz), 3.49 (1H, dd, J = 6.7, 9.7Hz), 3.62 (1H, dd, J = 7.0, 9.7Hz), 3.70 (1H, dd, J = 5.0, 9.7Hz), 4.02 (3H, m), 7.39 (6H, m), 7.67 (4H, m)

MSm / z283 (M ⁺ -tBu)

[Reference Example 6]

(2 S, 3 S)-1 (t-butyldiphenylsilyloxy) 2-methyl-6-trimethylsilyl-5-hexyn-3-ol (5

2) and (2S, 3R) 1 (t-butyldiphenylsilyllo) Xy) — 2 — Methyl — 6 — Trimethylsilyl — 5 — Hexin — 3 — Synthesis of all (53)

Under argon atmosphere, dissolve ethynyltrimethylsilane (780 ml, 5.0 mmo1) in 40 ml of THF, and add

(4.5 ml, 5.0 mmo 1) was added and stirred for 20 minutes. This was cooled to 178 ° C and the mixture (51) (1.7 g, 5.0 mm o

In addition to the TH F solution 4 0 m 1 of _{1), BF 3 'E t} 2 〇 (9. 5 ml, 5. added 0 mm o 1) and stirred for 15 minutes, the further from returning to room temperature 2 Stirred for hours. To this was added a saturated aqueous ammonium chloride solution, and the mixture was extracted with ethyl acetate. The ethyl acetate layer was washed with a saturated saline solution, dried over magnesium sulfate, and the solvent was distilled off. The crude product was purified by column chromatography (51 g, 2% Et ₂ O—hexane).

Both (52) (1.2 g, 52%) and (53) (1.1 g, 49%) were obtained as colorless oils.

(52)

¹ screen R (400MHz, CDCI3 / TMS) 6:

0.14 (9Η, s), 1.00 (3Η, d, J = 7.0Hz), 1.06 (9H, s), 1.92-1.99 (1H, m), 2.42 (1H, dd, J = 7.0, 10.1Hz ), 2.50 (1H, dd, J = 6.7, 10.1Hz), 2.84

(1H, d, J = 3.1Hz), 3.67 (1H, dd, J = 6.4, 10.2Hz), 3.75 (1H, dd, J = 4.2, 10.2Hz), 3.79 (1H, dd, J-4.3, 10.4 Hz), 7.37-7.46 (6H, m), 7.65-7.

68 (4H, m)

MSm / z381 (M ⁺ -tBu), 269 (M-Me-2Ph), 239 ( ⁺ -2Ph-3Me)

(53)

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

0.15 (9H, s), 0.91 (3H, d, J = 7.1 Hz), 1.07 (9H, s), 1.93- 1.99 (1 H, m), 2.46 (1H, dd, J = 6.4, 10.6 Hz), 2.54 (1H, dd, J = 6.4, 10.6Hz), 2.84 (1H, d, J = 3.1Hz), 3.67 (1H, dd, J = 6.4, 10.4Hz), 3.74-3.76 (1H, m ), 3.79 (1H, dd, J = 4.3, 10.4Hz), 7.37-7.46 (6H, m), 7.65-7.68 (4H, m) MSm / z423 (M ⁺ -e), 365 (M ⁺ -TS) ,

308 (M ⁺ -TMS-tBu)

[Reference Example 7]

Synthesis of MT PA ester (Determination of absolute structure of alcohol (X)) Under argon atmosphere, each of the above alcohols was dissolved in dry dichloromethane, and DMAP (2 equivalents), (R) — or (S) — MT PAC 1 (2 equivalents) was added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was purified as it was by TLC (10% AcOEt-hexane) to obtain an MTPA ester. Synthesis of Compound (54) (55) from Compound (52)

(52) (R) -MTPA ester (54)

(S) -MTPA ester (55)

(54) (R)

Yield: 30% (colorless oil) _{'HNMR (400MHz, CDC1 3 /} TS) <5:

0.12 (9H, s), 0.80 (3H, d, J = 6.7Hz), 1.06 (9H, s), 2.17 (1H, q, J = 6.7Hz), 2.68 (1H, t, J = 6.7Hz), 3.41 (2H, dd, J = 3.0, 10.3Hz), 3.58 (3H, s), 5.46 (1H, dd, J = 6.1, 10.3Hz), 7.28-7.46 (9H, m), 7.49 -7.55 (2H, m), 7.61-7.65 (4H, in)

(55) (S)

Yield: 25% (colorless oil)

_{'HNMR (400MHz, CDC1 3 /} TS) δ:

0.12 (9H, s), 0.86 (3H, d, J = 7.OHz) 1.07 (9H, s), 2.28 (1H, q, J = 6.1Hz), 2.57 (1H, dd, J = 5.8, 10.6 Hz), 2.71 (1H, dd, J = 6.1, 10.6Hz), 3.46 (3H, s), 3.48 (H, m), 5.49 (1H, dd, J = 5.8, 9.8Hz 7.28-7.46 (9H, m), 7.49-7.56 (3H, m), 7.60-7.69 (4H, m) Synthesis of Compounds (56) and (57) from Compound (53)

(53) (R) -MTPA ester (56)

(S) -MTPA ester (57)

(5 6) (R)

Yield: 13% (colorless oil)

_{'HNMR (400MHz, CDC1 3 /} TS) δ:

0.07 (9H, s), 0.95 (3H, d, J = 7.0Hz), 1.05 (9H, s), 2.26 (1H, q, J = 6.7Hz), 2.55 (1H, dd, J = 6.1) , 11.6Hz), 2.75 (1H, dd, J = 5.2, 11.6Hz), 3.42 (3H, s), 3.56 (1H, dd, J-5.8, 10.7Hz), 3.64 (1H, dd, J- 6.5, 10.7H z), 5.27 (1H, dd, J = 5.8, 11.6Hz), 7.28-7.45 (9H, m), 7.50-7.56 (2H, m), 7.59- 7.65 (4H, m)

(5 7) (S)

Yield: 17% (colorless oil)

1 删 R (400MHz, CDCI3 / TMS) δ:

0.11 (9Η, s), 0.82 (3Η, d, J = 7.OHz) 1.05 (9H, s), 2.19 (1H, q, J = 6.1Hz), 2. 58 (1H, dd, J = 6.7, 11.OHz), 2.75 (1H, dd, J = 6.7, 11.OHz), 3.

49 (1H, dd, J = 5.4, 10.3Hz), 3.54 (1H, dd, J = 5.8, 10.3Hz), 3.57 (3H, s), 5.32 (1H, dd , J = 6.7, 10.3Hz), 7.28-7.45 (9H, m), 7.59-7.54 (2H, m), 7.59-7.65 (4H, 1)

[Reference Example 8]

(4 R, 5 S) — 6 (t -butyldiphenylsilyloxy) 5 methyl-4 _ tetrahydrobilanyloxy — 1 — trimethylsilyl

—Synthesis of Hexin (58)

(53) (58) DHP (0.34 ml, 3.75 mm) was added to a solution of the alcohol (53) (1.07 g, 2.50 mmol) in dichloromethane (10 ml). o 1, 1.05 eq) and Ts〇H (72 mg, 0.375 mm o 1 _: 0.15 eq) were added and left overnight at room temperature. A saturated aqueous sodium hydrogen carbonate solution was added to the reaction solution, and the mixture was extracted with ethyl acetate. The extract was washed with brine, dried over magnesium sulfate, and the solvent was distilled off. The obtained crude product was subjected to silica gel column chromatography (100 g, l% Ac 〇 E t — Purification was performed to obtain (58) (1.26 g, 98%) as a colorless oil.

¹ face R (400MHz, CDCI3 / TMS) δ:

0.127 (9 / 2Η, s), 0.135 (9 / 2Η, s), 0.95 (3H, d, J = 7.0Hz), 1.058 (9 / 2H, s), 1.061 (9 / 2H, s) ), 1.41-1.62 (4H, m), 1.69-1.81 (2H, m), 2.09-2.17 (1H, m), 2.38 (1 / 2H, dd, J = 7.3, 17.1Hz), 2.46 ( 1 / 2H, dd, J = 4.6, '17.1Hz), 2.54 (1 / 2H, dd, J = 5.5, 17.1Hz), 2.66 (1 / 2H, dd, J = 5.8, 17.1Hz) , 3.38-3.49 (1H, m), 3.58-3.71 (2H, m), 3.75-3.81 (1H, m), 3.8 8-3.91 (1 / 2H, m), 3.92-4.06 (1 / 2H, m) , 4.66 (1 / 2H, dd, J = 3.1, 3.4Hz), 4.86 (1 / 2H, dd, J = 2.7, 4.3Hz), 7.35-7.44 (6H, m), 7.65-7.70 (4H, m)

[Reference Example 9]

(2S, 3R) _2-Methyl_3-tetrahydrobilanyloxy

Synthesis of 5-hexin-1-ol (59)

匕 F solution (20 ml) of 匕合合 (58) (1.13 g, 2.20 mm ο 1) IM nBu ₄ NF / T F (8.8 ml, 8.8 Ommol, 4 equivalents) was added and the mixture was stirred at room temperature for 4 hours. Water was added to the reaction solution, which was extracted with ethyl acetate. The extract was washed with brine, dried over magnesium sulfate, and the solvent was distilled off. The obtained crude product was subjected to silica gel column chromatography (35 g, 20% A c 〇 E t — Purification was performed to obtain colorless oily (59) (450 mg, 96%).

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

0.99 (3 / 2Η, d, J = 6.7Hz), 1.01 (3 / 2H, d, J = 7.0Hz), 1.41-1.89 (6H + 1 / 2H, m), 1.99 (1 / 2H, t, J = 2.7Hz), 2.00 (1 / 2H, t, J = 2.7Hz), 2.13-2.19 (1 / 2H, m), 2.33 (1 / 2H, bs), 2.38 (1 / 2H, ddd, J = 2.4, 6.1, 17.1Hz), 2.57 (1 / 2H, ddd, J = 2.4, 4.0, 17.1Hz), 2.63 (1 / 2H, ddd, J = 2.8, 4.0, 17.1Hz), 2.72 (1 / 2H, ddd, J = 2.8, 7.0, 17.1Hz), 3.30-3.31 (1 / 2H, m), 3.41-3.56 (3 / 2H, m), 3.60-3.81 (2H, m), 3.95-4.01 (3 / 2H , m), 4.69-4.71 (1H, m)

[Reference Example 10]

Synthesis of (4 R, 5 R) 1-3 Hydroxy-1- 4-methyl-5-tetrahydrobilanyloxy 1-octen-1-yne (61)

(59) (60) (61)

Oxalyl chloride (0.56 ml, 6.30 mmo 1, 3 eq.) Was added to dichloromethane solution (4 ml) of DMSO (0.92 ml, 12.5 mmo 1, 6 eq.). In addition, the mixture was stirred at −78 ° C. for 1 hour under an argon atmosphere. A solution of compound (59) (44 O mg, 2.08 mmo 1) in dichloromethane (10 ml) was added to the resulting solution at −78 ° C., and the mixture was stirred for 30 minutes. ₃ N (3.2 ml, 24 mmo 1, 12 equivalents) was added, and the mixture was stirred at —78-0 ° C. for 1 hour. Water was added to the reaction solution, which was extracted with ethyl acetate. Wash the extract with saturated saline, and add magnesium sulfate After drying with a solvent, the solvent was distilled off. The crude product was filtered through a small amount of silica gel, using a the next reaction without solvent evaporated to a colorless oil of aldehyde (6 0) The resulting _t The product further purification. To a solution of aldehyde (60) (426 mg, 2.02 mmo 1) in THF (10 ml) at 0 ° C was added 1 M biel magnesium bromide THF (4.0 ml, 4.00 ml). mmo1, 2 equivalents) and stirred at 0 ° C. for 1 hour. Water was added to the reaction solution, which was extracted with ethyl acetate. The extract was washed with saturated brine, dried over magnesium sulfate, the solvent know _c obtained crude product was distilled off force Gerukaramuku port Matogurafi (5 5 g, 2 0% A c 0 E t - hexane) Then, colorless oily aryl alcohol (61) (329 mg, 68%) was obtained.

'HNR (400MHz, CDC1 TMS) δ:

0.85 (3 / 4Η, d, J = 7.OHz), 0.88 (3 / 4Η, d, J = 7.3Hz), 0.90 (3 / 4H, d, J2 7.0Hz) , 0.93 (3 / 4H, d, J = 7.OHz), 1.47-1.87 (6H, m), 1.98-2.05 (1H, m), 2.15-2.19 (1H, m), 2.37-2.89 (2H, m), 3.37-4.15 (4.5 H, in), 4.51-4.84 (1.5H, m), 5.13-5.35 (2H, m), 5.83-5.94 (1H, m)

[Reference Example 11]

(3 R, 4 R, 5 R) — 3, 5 — dihydroxy— 4-methyl— 1 — octene — 7 — yne (22) and (3 S, 4 R, 5 R)-3, 5 — dihydroxy Synthesis of 4-methyl-1- 1-octene 7-yne (23)

TsOH (25 mg, 0.13 mmo 1: 0.1 equivalent) in 10 ml of methanol solution of aryl alcohol (61) (315 mg, 1.32 mmol) Was added and left at room temperature for 1 hour. Saturated aqueous sodium bicarbonate was added to the reaction mixture, and the mixture was extracted with E t ₂ 〇. The extract was washed with saturated saline and dried over magnesium sulfate, and the solvent was distilled off. The obtained crude product was purified by silica gel column chromatography (55 g, 10% Ac〇Et-hexane) to obtain a colorless oily ene-in compound (22) (79 mg). , 39%) and (23) (75 mg, 37%).

( twenty two )

'HNMR (400MHz, CDCU / TMS) δ

0.90 (3Η, d, J = 7.0Hz), 1.95 (1H, dquin, J = 2.8, 7.0Hz), 2.08 (1H, t, J = 2.8Hz), 2.43 (1H, ddd, J-2.8, 7.0, 17.1Hz), 2.54 (1H, ddd, J = 2.8,

4.6, 17.1Hz), 2.72 (1H, d, J = 5.5Hz), 2.96 (1H, d, J = 4.6Hz), 3.79 (1H, tt, J = 4.6, 7.0Hz), 4.44 (1H, dt t, J = 7.0, 1.5, 5.5Hz), 5.23 (1H, dt, J = 10.7, 1.5Hz), 5.32 (1H, dt, J = 17.1, 1.5Hz), 5.94 (1H.ddd, J =

(5.5, 10.7, 17.1Hz)

( twenty three )

'HNMR (400MHz, CDCU / TMS) <5

0.83 (3H, d, J = 7.0Hz), 1.83 (1H, dquin, J = 7.0, 7.9Hz), 2.07 (1H, t, J = 2.8Hz), 2.41 (1H, ddd, J = 2.8, 6.7, 16.8Hz), 2.58 (1H, ddd, J = 2.8, 4.0, 16.8Hz), 2.88 (1H, bs), 3.41 (1H, bs), 3.74 (1H, m), 4.14 (1H, 11, J = l .2, 7.3Hz), 5.19 (1H, dt, J = 10.4, 1.2Hz), 5.27 (1H, dt, J = 17.1, 1.2Hz), 5.88 (1H, ddd, J = 7.3, 10.4, 17.1 Hz)

'[Reference Example 1 2]

Synthesis of (3R, 4R, 5R) -3,5-Di (tert-butyldimethylsilyloxy) -4-methyl-1-octene-7-yne (38)

(38)

2,6-Lutidine (0.18 ml, 1.5 mmol, 4 equivalents) was added to a dichloromethane solution (5 ml) of the compound (22) (58 mg, 0.376 mmo 1). Then, TBS〇Tf (0.34 ml, 1.5 mmol, 4 equivalents) was added, and the mixture was stirred at 0 ° C. for 1 hour. Saturated aqueous sodium hydrogen carbonate was added to the reaction mixture, and the mixture was extracted with ethyl acetate. The extract was washed with brine, dried over magnesium sulfate, and the solvent was distilled off. The obtained crude product was purified by silica gel column chromatography (10 g, 2% AcOEt-hexane) to obtain colorless oil (38) (141 mg, 98%). Was.

¹ stroke R (400MHz, CDCI3 / TMS) δ:

0.01 (3Η, s), 0.5 (3Η, s), 0.07 (3H, s), 0.11 (3H, s), 0.89 (9H, s), 0.90 (9H, s) ), 0.90 (3H, d, J = 7.0Hz), 1.78 (1H, dquin, J = 4.9, 7.0Hz), 1.93 (1H, t, J = 2.8Hz), 2.26 (1H, ddd, J-2. 8, 7.0, 16.8 Hz), 2.40 (1H, d dd, J = 2. 8, 4. 3, 16.8 Hz), 3.86 (1H , dt J = 7.0, 4.3 Hz), 4.11 (1H, ddt, J = 5.8, 7.3, 1.8 Hz), 5.09 (1H, dt, J = 10.1, 1.8Hz), 5. 14 (1H, dt, J = 17.4, 1.8 Hz), 5.84 (1H, ddd, J = 7.3, 10.1, 17.4Hz)

[Reference Example 13]

Synthesis of acetonide (Determination of absolute configuration of en-yne compound (II)) The above-mentioned en-yne compound (5 mg) was dissolved in 0.4 ml of acetone, and 0.1 ml of dimethoxypropane was added. CSA (1.5 mg, 0.2 equivalent) was added, and the mixture was left at room temperature for 5 hours. The solvent is distilled off to obtain The obtained crude product was purified by silica gel column chromatography (6 g, 5% AcOEt-hexane) to obtain acetonide.

Synthesis of Compound (62) from Compound (22)

Yield: 80% (colorless oil)

1 recitation R (400MHz, CDClj / TMS) δ:

0.90 (3Η, d, J = 7.0Hz), 1.39 (3H, s), 1.40 (3H, s), 1.86- 1.92 (1H, m), 2.01 (1H, t, J = 2.8Hz), 2.44 ( 1H, ddd, J = 2.8, 6.1, 17.4Hz), 2.48 (1H, ddd, J = 2.8, 5.5 '17.4Hz), 3.49 (1H, dt, J = 7.6, 5.8Hz), 4.43 (1H , ddt, J = 6.1, 5.2, 1.5Hz), 5.17 (1H, dt, J = 10.7, 1.2Hz), 5.26 (1H, dt, J = 17.4, 1.2Hz), 5.79 (1H, ddd, J = 6.1, 10.7, 17.4Hz)

^{_{1 3 CNMR (lOOMHz, CDC1 3}} / TMS) δ:

12.89 (d), 24.10 (d), 25.24 (q), 29.70 (t), 39.76 (d), 69.66 (s), 70.61 (d), 73.02 (d), 80.96 (d), 100.88 (s), 115.77 (t), 135.59 (t)

Synthesis of Compound (63) from Compound (23)

Yield: 80% (colorless oil) _{'HNMR (400MHz, CDC1 3 /} TMS) δ:

0.82 (3H, d, J = 6.7Hz), 1.45 (3H, s), 1.49 (3H, s), 1.51-1.61 (1H, m), 2.01 (1H, t, J-2.7Hz), 2.42 (1H , ddd, J = 2.7, 5.5, 17.4 Hz), 2.52 (1 H, ddd, J = 2.7, 4.0, 17.4 Hz), 3.68 (1H, ddd, J = 4.0, 5.8, 10.1 Hz), 3.91 (1H , ddt, J = 7.3, 10.1, 1.5Hz), 5.24 (1H, dd, J = l.5, 7.3Hz), 5.29 (1H, dd, J = l.5, 17.4Hz), 5.76 (1H, ddd, J = 7.3, 10.1, 17.4Hz), ^13C band R (100MHz, CDClg / TMS) 6:

12.15 (a), 19.71 (q), 29.70 (t), 30.04 (q), 39.76 (d), 69.66 (s), 70.61 (d), 73.02 (d), 80.96 (d), 100.88 (s ), 115.77 (t), 135.59 (t) The following enyne compounds were synthesized by using a suitable raw material and applying a similar production method.

[Reference Example 14]

Synthesis of (3S, 4R, 5R) _3,5-bis (t-butyldimethylsilyloxy) —4-methyl-1-octen-17-yne (39)

(39)

'HNMR (400MHz, CDCU / TMS) <5

0.02 (3H, s), 0.057 (3H, s), 0.063 (3H, s), 0.11 (3H, s), 0.78 (3H, d, J = 7.0Hz), 0.86 (9H, s), 0.90 (9H, s), 1.89 (1H, dquin, J = 5.5, 7.OHz), 1.93 (1H, t, J = 2.8 Hz), 2.26 (1H, ddd, J = 2.8, 7.0, 16.8 Hz), 2.3 9 (1H, ddd, J = 2.8, 4.0, 16.8Hz), 3.97 (1H, ddd, J = 4.0, 5.2, 6.7Hz), 4.12 (1H, ddt, J = 6.4, 6.7, 1.2Hz), 5.09 (1H, dt, J = 10.4, 1.2Hz), 5.16 (1H, dt, J-17.1, 1.2Hz), 5.75 (1H, ddd, 6.1, 10.4, .17.1 Hz) Sm / z382 ( ⁺ ), 367 (M Me), 325 (M ⁺ -tBu)

[Reference Example 15]

(3 R, 4 R, 5 S) Synthesis of 3,5-bis (t-proxy) —4-methyl-1-octene_7-yne (40)

(40)

_{• HNMR (400MHz, CDC1 3 /} TS) <5:

0.01 (3H, s), 0.049 (3H, s), 0.051 (3Η, s), 0.08 (3H, s), 0.89 (18H, s), 0.92 (3H, d, J = 7.0Hz), 1.86 (1H , dquin, J = 4.0, 6.7Hz), 1.95 (1H, t, J = 2.8Hz), 2.38 (2H, dd, J = 2.7, 5.8Hz), 3.88 (1H, ddd, J = 4.0, 6.1) , 6.4Hz), 4.09 (1H, t, 7.0), 5.10 (1H, dt, J = 10: 4, 1.5Hz), 5.14 (1H, dt, J = 17.4, 1.5Hz), 5.81 (1H, ddd, J = 7.0, 10.4, 17.4)

^{Sm / z382 (M +),} 367 (M + - e), 325 (M + -tBu)

[Reference Example 16]

Synthesis of (3 S, 4 R, 5 S) 3,5-bis (t-butyldimethylsilyloxy) — 4-methyl-1-octen-7-yne (41)

(41) _{'HNMR (400MHz, CDC1 3 /} TS) δ

0.03 (3H, s), 0.06 (3H, s), 0.07 (3H, s), 0.08 (3H, s), 0.76 (3H, d, J: 7.0Hz), 0.889 (9H, s), 0.892 (9H , s), 1.91 (IH, dquin, J = 3.7, 7.OH z), 1.97 (IH, t, J = 2.8 Hz), 2.36-2.40 (2H, m), 3.99-4.05 (2H, m), 5.09 (IH, dt, J = 10.4, 0.9Hz), 5.13 (IH, dt, J = 17.1, 0.9Hz), 5.73 (IH, ddd, J = 7.6, 10.1, 17.1Hz) )

Sm / z382 (M ⁺ ), 367 (M ⁺ -Me), 325 (M ⁺ -tBu)

[Reference Example 17]

(3 R, 4 S, 5 R) 3, 5-bis (t-butyldimethylsilyl)

Synthesis of 4-methyl-1-octene-7-yne (4 2)

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

0.03 (3Η, s), 0.06 (3H, s), 0.07 (3H, s), 0.08 (3H, s), 0.76 (3H, d, J = 7.0Hz), 0.889 (9H, s), 0.891 (9H , s), 1.91 (IH, dquin, J = 3.7, 7.OH z), 1.97 (IH, t, J = 2.8 Hz), 2.31-2.43 (2H, i), 3.98-4.04 (2H, m), 5.10 (IH, dt, J = 10.1, 1.5Hz), 5.13 (IH, dt, J = 17.1, 1.5Hz), 5.74 (IH, ddd, J = 7.6, 10.1, 17.1Hz)

MSm / z382 (M ⁺ ), 367 (MMe), 325 (M ⁺ -tBu)

[Reference Example 18]

(3 S, 4 S, 5 R) — Synthesis of 3,5 —bis (t—bryloxy) 4 methyl— 1 —octen-1 7 —yne (43)

_{'HNMR (400MHz, CDC1 3 /} TMS) δ:

0.01 (3Η ', s), 0.049 (3H, s), 0.051 (3H, s), 0.08 (3H, s), 0.89 (18H, s), 0.92 (3H, d, J = 7.0Hz), 1.85 ( IH, dquin, J = 3.7, 6.7Hz), 1.96 (IH, t, J = 2.8Hz), 2.39 (2H, dd, J-2.8, 6.7Hz), 3.88 (IH, ddd, J = 4.0, 6. 1, 6.4Hz), 4.07 (IH, t, J = 6.7Hz), 5.10 (IH, dt, J = 10.1, 1.8Hz) .5.14 (IH, dt, J = 18.3, 1.8Hz) ), 5.81 (IH, ddd, J = 7.0, 10.4, 17.4Hz) MSm / z 367 (M ⁺ -Me), 325 (M ⁺ -tBu) [Reference Example 19]

(3R, 4S, 5S) Synthesis of 3,5_bis (t-bryloxy) -14-methyl-_1-octen-7_yne (44)

_{'HNMR (400MHz, CDC1 3 /} TMS) δ

0.01 (3H, s), 0.057 (3H, s), 0.063 (3H, s), 0.11 (3H, s), 0.78 (3H, d, 7.0Hz), 0.86 (9H, s), 0.90 (9H , s), 1.88 (IH, dquin, J = 5.5, 6.7 Hz), 1.93 (IH, t, J = 2.8 Hz), 2.26 (IH, ddd, J = 2.8, 7.0, 16.8 Hz), 2.3 9 ( IH, ddd, J = 2.8, 4.0, 16.8Hz), 3.97 (IH, dt, J = 4.0, 5.5Hz), 4.12 (1H, ddt, J = 5.2, 6.7, 1.2Hz), 5.09 (1H, dt, J = 10.4, 1.2Hz), 5.15 (1H, dt, J = 17.1, 1.2Hz), 5.75 (1H , ddd, J = 6.7, 10.4, 17.4Hz) MSm / z382 (M ⁺ ), 367 (MMe), 325 (M-tBu) [Reference Example 20]

(3 S, 4 S, 5 S) Synthesis of 3, 5 —bis (t-broxy) — ₄ —methyl— 1 —octen-1 7 —yne (45)

¹ stroke R (400MHz, CDCI3 / T1 S) δ:

0.01 (3Η, s), 0.05 (3Η, s), 0.07 (3H, s), 0.10 (3H, s), 0.88 (3H, d, J = 7.0Hz), 0.89 (9H, s), 0.90 (9H, s), 1.76-1.80 (1H, m), 1.93 (1H, t, J = 2.8 Hz), 2.26 (1H, ddd, J = 2.7, 7.0, 16.8 Hz), 2.40 (1H, ddd, J = 2.7,

4.3, 16.8Hz), 3.85 (1H, dt, J = 7.0, 4.3Hz), 4.11 (1H, ddt, J = 5.8, 7.3, 1.8Hz), 5.10 (1H, dt, J = 10.1, 1.8Hz) ), 5.14 (1H, dt, J = 17.4, 1.8 Hz), 5.84 (1H, ddd, J = 7.3, 10.1, 17.4Hz)

MSm / z382 (M ⁺ ), 367 (M ⁺ -Me), 325 (M ⁺ -tBu)

[Example 1]

(2 OS) _ 1, 25-dihydroxy-1-2-methyl-1 3 j3-synthesis of bimin D ₃ (72)

The exomethylene compound (92) (17 mg) was dissolved in 0.3 ml of toluene, and Et ₃ N (0.45 ml) was added under an argon atmosphere. _{P d 2 (dba) 3 ·} CHC 1 3 (1. 9 mg, 0 - 0 3 _{eq), P h 3 P (2.} 5 mg, 0. 3 eq) was added and stirred at room temperature, then E A solution of the quinine compound (42) (13 mg, 0.7 equivalents) in toluene (0.2 ml) was added, the mixture was stirred at room temperature for 10 minutes, and further placed in an oil bath at 120 ° C. 2. The reaction was performed for 5 hours. After cooling, the reaction solution was filtered and purified by silica gel chromatography (AcOEt: hexane = 1: 3) to obtain compound (80).

The obtained compound (80) was dissolved in 1 ml of methanol, added with CSA (ll mg, 1 equivalent), and allowed to react overnight at room temperature under an argon atmosphere. The reaction solution was evaporated, purified water was added, and the mixture was extracted with AcOEt. The extract was washed with saturated saline, dried over magnesium sulfate, and the solvent was distilled off. The resulting crude product was purified by silica gel chromatography (A c 0 E t: hexane = 1: 1) and recrystallized with recycling preparative HPLC (L ichrosorb RP - 1 8, 7 0% M e CN / H 2 O ) To give colorless crystals (72) (9.3 mg, 63%).

HNR (400MHz, CDC1 ₃ -D20 / TMS) 6 0.53 (3H, s), 0.85 (3H, d, J = 6.7Hz), 1.08 (3H, d, J = 6.8Hz), 1.21 (6H, s), 1.12-2.04, (19H, m) 2.23 (1H , dd, J = 7.9Hz, 13.4Hz), 2.67 (1H, dd, J = 4.0Hz, 13.4Hz), 2.83 (1H, in), 3.83 (1H, td, 1 = 7.9Hz, 4.0Hz), 4.29 (1H, d, 1 = 3.3Hz), 5.01 (1H, d, J = l.8Hz), 5.28 (1H, m), 6.01 (1H, d, J-ll.3Hz), 6.3 9 (1H, d , J = l 1.3Hz)

UV (EtOH): λ max266nm

^{MS m / z 430 (+)} , 412 (M + -H 2 0), 394 (M + -2H 2 0)

HR-MS:. Calcd for C 28 H 46 0 3: 430.3447

found: 430.3443 The following 1,25-dihydroxy-2-methylvitamin D ₃ derivative was produced by using the same reaction conditions as in Example 1.

[Example 2]

(20 S) — 1, _25-dihydroxy -2) 3-methyl-3; 8-synthesis of vitamin 1 _Ί ( ₆₈ )

Ή-NMR (400MHz, CDC 1 3 -D 2 0 / TMS) δ:

0.55 (3Η, s), 0.85 (3H, d, J = 6.4Hz), 1.15 (3H, d, J = 6.7Hz), 1.21 (6H, s), 1.17-2.01 (19H, m) 2.42 (1H, dd, J = 13.9, 4.9Hz), 2.52 (1H, d, J = 13.9Hz) 2.82 (1H, dd, J = ll.9Hz, 4.0Hz), 3.99- 4.04 (1H + 1H, m), 5.02 ( 1H, t, Jl. 8Hz), 5.37 (1H, t, J = l.8Hz), 6.03 (1H, d, J = l 1.3Hz), 6.35 (1H, d, J = l 1.3 Hz)

UV (EtOH): λmax263nm

^{MS m / z 430 (M +} ), 412 (M + -H 2 0), 394 (+ -2H 2 0)

HR-MS:. Calcd for C 28 H 46 0 3: 430. 3447

found: 430. 3441

[Example 3]

(2 O S) — 1 β, 25 — Dihydroxy — 2; 3 — Methyl-3 | 3 — Synthesis of biquinone D (69)

(69)

Ή-NMR (400MHz, CDC1 TMS) δ

0.55 (3H, s), 0.85 (3H, d, J = 6.7 Hz), 1.22 (6H, s), 1.23 (3H, d, J = 7.3 Hz), 2. 17 (1H, d, J = 4.3Hz), 2.50 (1H, brd, J = 12.5Hz), 2.59 (1H, dd, J = 14.0Hz, 3.7Hz), 2.79 (1H, d, J = 7.6Hz), 2.85 (1H, dd, J = 12.5Hz, 4, 9Hz), 3.91 (1H, m), 4.17 (1H, HI), 5.01 (1H, d, J = 2.1Hz), 5.25 (1H, d, J = l. 8Hz), 6.09 (IH, d, J = ll.3Hz), 6.48 (IH, d, 11.3Hz) )

MS m / z 430 (M, 412 (M and H ₂₀ ), 394 ( ⁺ -2H ₂₀ ), 379 (M 2H ₂₀ -Me)

HR-MS: calcd. For C ₂₈ H ₄₆ 0 ₃ ; 430.3447, found; 430.3446 [Example 4]

Synthesis of (20S) -1α, 25-dihydroxy 2 / 3-methyl-3-hydroxy-bi-min D ₃ (70)

_{Ή-NMR (400MHz, CDC1 3} / TMS) δ:

0.54 (3Η, s), 0.85 (3Η, d, J = 6.4Hz), 1.06 (3H, d, J = 7.0Hz), 1.22 (6H, s), 2.12 (IH, d, J = 2.8Hz), 2.34 (IH, dd, 14.7Hz, 7.0Hz), 2.60 (IH, brs), 2.64 (IH, dd, J = 13.4Hz, 2.8Hz), 2.84 (IH, dd, J = ll.6Hz, 3.1Hz), 3.65 (IH, m), 3.90 (IH, m), 5.05 (IH, d, J = l.8 Hz), 5.30 (IH, d, J = 2.7 Hz), 6.02 (IH, d, J = ll.3Hz), 6.41 (IH, d, 11.3Hz)

MS m / z 430 (M ^{_{ten), 412 (+ -H 2 0}} ), 394 (M + -2H 2 0), 379 (M + -2H 2 0- e)

HR-MS: calcd. For C ₂₈ H ₄₆ 0 ₃ ; 430.3447, found; 430.3447

[Example 5]

(20 S) — I j3, 25-Dihydroxy 2/3-Methyl-3 a-Biminine Synthesis of D 3 (71)

(71) 'H-NMR ( 400MHz, CDC1 3 / TMS) <5:

0.54 (3H, s), 0.85 (3H, d, J = 6.4 Hz), 1.10 (3H, d, J = 6.7 Hz), 1.22 (6H, s), 1.68 (2H, m), 1.85 (2H, m ), 1.98 (2H, m), 2.24 (1H, dd, J = 13.4Hz, 8.5Hz), 2.65 (1H, dd, J = 13.4Hz, 4.3Hz), 2.82 (1H, dd, J = 12.2Hz, 4.3Hz), 3.81 (1 H, m), 4.27 (1H, m), 5.02 (1H, d, J = 2.1 Hz), 5.28 (1H, d, J = l.8 Hz), 6.02 (1 H, d , J = ll.3Hz), 6.40, (1H, d, J = l 1.3Hz)

MS m / z 430 (M + ), 412 (M + -H 2 0), 394 (M + -2H 2 0), 379 (M tens - 2H ₂ 0- Me)

HR- S: calcd. For C ₂₈ H ₄₆ 0 ₃ ; 430.3447, found; 430.3446

[Example 6]

(20 S) — 1) 3,25 —Dihydroxy—2H-Methyl-1 / 3 / 3-Biminemin D Synthesis of D (73)

_{Ή-NR (400MHz, CDC1 3} / TS) δ:

0.55 (3H, s), 0.85 (3H, d, J = 6.4 Hz), 1.02 (3H, d, J = 7.OHz), 1.2 (6H, s), 1. 83 (IH, m), 2.00 (2H, m), 2.11 (IH, m), 2.27 (IH, d, J = 7.OHz), 2.34 (IH, dd, 1 = 14. OHz, 5.5Hz), 2.65 (IH, dd, J = 14.OHz, 7.8Hz), 2.84 (IH, dd, J = l 2.2Hz, 4.3Hz), 3.72 (IH , m), 3.97 (IH, t, J = 4.9 Hz), 5.07 (IH.d, J = 2.IH z), 5.30 (IH, d, J = 2.1 Hz), 6.04 (IH, d, J = l 1.3 Hz), 6.43 (IH, d, J = ll. 3H z)

MS m / z 430 (M + ), 412 (M + -H 2 0), 394 (M + -2H 2 0), 379 (M + -2H 2 0-Me)

HR-MS: calcd. For C ₂₈ H ₄₆ 0 ₃ ; 430.3447, found; 430.3445

[Example 7]

(20S) -l, 25—Synthesis of dihydroxy-1-o! —Methyl-3-bi-yubmin D (74)

_{Ή-NMR (400MHz, CDC1 3} / TMS) δ:

0.53 (3Η, s), 0.85 (3Η, d, J = 6.4 Hz), 1.21 (6H, s), 1.22 (3H, d, J = 7.OHz), 2.09 (IH, d , J = 4.6Hz), 2.49 (IH, d, J = 14.7Hz), 2.58 (IH, dd, J = 14.OHz, 3.7Hz), 2.80 (IH, d, J = 7.9Hz), 2.85 (IH, m), 3.91 (1H, m), 4.17 (IH, m), 4.98 (IH, d, J = 2.1Hz), 5.23 (IH, d, J = l. 8Hz), 6.03 (IH, d, J = ll. 3Hz), 6.48 (IH, d, J = ll. 3Hz) ^{MS m / z 430 (M +} ), 412 (M + -H 2 0), 394 (+ -2H 2 0), 379 (+ -2H 2 0-Me)

HR-MS: calcd. For C ₂₈ H ₄₆ 0 ₃ ; 430.3447, found; 430.3447

[Example 8]

Synthesis of (20S) 1-1 / 3,25-dihydroxy-2α-methyl-3α-biamine D 2 (75)

_{Ή-NMR (400MHz, CDC1 3} / TMS) δ:

0.53 (3Η, s), 0.85 (3Η, d, J = 6.4Hz), 1.13 (3H, d, J = 6.7Hz), 1.21 (6H, s), 1.69 (2H, m), 1.84 (2H, m ), 1.98 (2H, m), 2.41 (1H, dd, J = 13, 7Hz, 5.5Hz), 2.51 (1H, dd, J = 13.4Hz, 2.4Hz), 2.82 (1H, m), 4.02 -4.08 (2H, m), o.01 (1H, d, Jl.8Hz), 5.35 (1H, d, J = l.8Hz), 6.01 (1H, d, J = l 1.6Hz), 6.36 ( (1H, d, J = l 1.6Hz)

^{MS m / z 430 (M +} ), 412 (M + -H 2 0), 394 (M + -2H 2 0), 379 (M + -2H 2 0-Me)

. HR-MS: calcd for C 28 H 46 0 3; 430.3447, found; 430.3445 [ Example 9]

結合 Binding affinity of the compounds of the present invention to thymus 1 and 25-dihydroxyvitamin D 3 receptor (VDR) One sample (approximately 25 mg) of thymus vitamin D receptor kit from Yamasa Shoyu Co., Ltd. in 0.05 M phosphoric acid 0.5 M potassium buffer (H7.4) 5.5 m1 Was dissolved. After an ethanol solution 5 0 1 and receptions evening first solution 5 0 0 1 of the test compound was 1 hour Pureinkyu Pies at room temperature, [2 6, 2 7 - methyl - ³ H] 1 a, 2 5 - dihydric de Rokishibitamin D _Three solutions 50 ^ 1 (13CiZmmol, 16, OOO dpm) were added to a final concentration of 0.1 nM, and incubated at 4 ° C for 1 hour. Bonded and unbonded [26, 27-Methyl- ³

H] 1 a, 2 5 - dihydric mud carboxymethyl Vitamin D ₃ is added to 2 0 0 1 dextran over Cote dough Chiya call centrifuged, 5 0 0 1 of the supernatant liquid Shinchireshiyo emissions cocktail (ACS- II) 9.5 ml was added, and the radioactivity was measured with a liquid scintillation counter.

Binding affinity for D ₃ receptions evening one test compound (VD R) is a dihydrazide mud carboxymethyl concentration which inhibits 50% binding of vitamin _{D 3 - [2 6, 2} 7 - ^methyl-3 H] 1, 2 5 It was calculated and expressed as a relative intensity ratio when 1 / 25-dihydroxyvitamin D ₃ was set to 100. The results are shown in the following table. Compound VDR binding affinity Compound VDR binding affinity la, 25- (0H) ₂ VD ₃ 100

Compound (65) 13 Compound (68) 160 Compound (1) 0.05 Compound (69) 0.03 Compound (2) 0.3 Compound (70) 0.08 Compound (3) 0.8 Compound (71) 7 Compound (4) 400 Compound (72) 1200 Compound (5) 0.05 Compound (73) 0.05 Compound (6) 4 Compound (74) 17 Compound (7) 0.06 Compound (75) 0.03 Here, the compounds (1) to (7) and the compound (65) in the table are comparative examples.

(20 R) — 1 β, 25 — dihydroxy 2-monomethyl — 3/3 — bimin D ₃ ,

(2 0 R) - 1 shed, 2 5 - dihydric de Proxy one 2) 3 - methyl one 3 Facial velvetleaf evening Min D _3,

(2 OR) — 1 β, 25 — dihydroxy — 2 j3 — methyl— 3 α— bi-min D ₃ ,

(2 OR) — 1 α, 25 — dihydroxy-1- 2 methyl 3) 3 — bimin D ₃ ,

(20 R) 1 1 β, 25 — dihydroxy — 2 α; — methyl — 3/3 — biminmin D ₃ ,

(2 0 R)-1 α, 25-dihydroxy-2α-methyl-3α-biminmin D ₃ ,

(2 0 R) - 1 β , 2 5 - dihydric Dorokishi 2 alpha - methyl - 3 Hibi evening Min D ₃ is

(2 0 R) - 1 shed, 2 5 - dihydric de proxy one 2 i3 - methyl-3/3 - bi evening Min D _3. [Example 10]

Effect of the compound of the present invention on the differentiation inducing action of HL-60 cells

The HL-60 cells used were purchased from a cell bank (Japanese Cancer Research Resource Bank, cell number: JCRB085). Cells were used as cryopreservation stocks to prevent changes in cell characteristics due to subculture, and those that had been thawed before starting the experiment and subculture started. The experiments were performed for one month to six months. For subculture, collect cells in suspension culture by centrifugation and add 1 Z to fresh culture solution.

1 0 0 about - was carried out in Rukoto it is diluted to a concentration of ^{(1 2 X 1 0 5 cells} / m 1). RPMI-164 medium containing 10% fetal bovine serum was used as a culture solution. The cells that had been subcultured were collected by centrifugation, dispersed in a culture solution at 2 × 10 ⁴ cells / m 1, and inoculated in a 24-well culture dish at 1 ml 1 Z-well. To the system was added an ethanol solution of the present invention compounds ^{(1 X 0- 9 M- 1 X} 1 0- 6 M) in Ueru per Ri 1 1. Note that one shed, for _{_{2 5 (OH) 2 D 3}} , was added a 1 X 1 0- ⁷ M to 1 X 1 0- ⁴ M ethanol solution Ueru per 1, the Etano Ichiru to control Added at 1 1 per well. After culturing at 37 ° C under 5% CO ₂ for 4 days, the cells were collected centrifugally. The measurement of the reduction activity of nitro blue tetrazolium (hereinafter NBT) was performed according to the following procedure. That is, the cells collected by centrifugation are suspended in a fresh culture solution, and then added so that 0.1% of NBT and 12-O-tetradecanoylphorpol-13-acetate 10OnM are obtained. After incubation at 37 ° C for 25 minutes, a cytospin specimen was prepared. After air-drying, the cells were stained with Kernehichroth and the ratio of cells positive for NBT reduction activity was determined under a light microscope. The results are shown in the following table.

Effect of the compound of the present invention on nitroble-reducing activity in HL-60 cells

Industrial applications

1 represented by the above formula (I) provided by the present invention, 2 5 _ dihydric Dorokishi _ 2 - methyl vitamin D ₃ derivative, diseases usefulness vitamin ₃ derivatives have been widely recognized (osteoporosis, rickets , Hyperparathyroidism, etc.). Among them, the compound of the present invention can be used particularly effectively for diseases (cellular cancer, psoriasis, etc.) caused by insufficient cell differentiation due to the extremely strong differentiation-inducing action.

Also, depending on the type of stereoisomer derived from the 1-, 2-, and 3-positions, certain isomers have high affinity for vitamin D receptor and also for vitamin D binding protein, The isomers show high affinity for the vitamin D receptor and low affinity for the vitamin D binding protein, indicating differences in affinity for both proteins. It can be used as a therapeutic.

Claims

Range of request

1. The following general formula (I)

[Wherein, R and R ₂ each independently represent a hydrogen atom or a tri (C i -C ₇ alkyl) silyl group. Here, the configurations of the asymmetric carbons at the 1-, 2-, and 3-positions are each independently para-coordinate or 13-coordinate. ]

A 1,25-dihydroxy-2-methylvitamin D ₃ derivative represented by the following formula:

2. The following general formula (II)

[In the formula, X represents a bromine atom or an iodine atom. ]

An exomethylene compound represented by the following general formula:

Claim 1 characterized by reacting an enyne compound represented by the following formula in the presence of a palladium catalyst and, if necessary, deprotecting a tri (C ^ -C hydrocarbon) silyl group. The method for producing a vitamin D ₃ derivative according to the above.